JOURNAL OF CHEMICAL 2 (1), JANUARY 2008: 40-44

ISOLATION AND IDENTIFICATION OF LEAF ANDONG steroida

glycoside ( Cordyline terminalis Kunth)

NW Bogoriani

Department of Chemistry, State University of Udayana, Bukit Jimbaran

ABSTRACT

Isolation and identification of two steroid glicosides have been conducted from Andong leaves ( Cordyline
terminalis Kunth). The isolates (4.0 mg of white amorphous solid) was Obtained after a series of chromatographic
separations. Identification of the isolates using mass spectrometry with electrospray positive Showed 868 MW as
calculated from the ion peaks of m / z 891 [M + Na] +, and 869 [M + H] +. The ion peaks of isolates at m / z 727 [(M + Na)
- 164] +, 723 [(M + H) - 146] +, 705 [(M + H) - 164] +, and 413 [(M + H) - 456] + of its fragments indicated resources to the
presence of three sugars (two sugars and one central terminal sugar) from methylpentose moeity (each of 164 MW) linked
to an agyicone. Proton magnetic resonance spectrum of the isolate in pyridine-d 5 Showed characteristic proton signals
for three steroid methyls (two angular methyls and one secondary methyl) at δ 1:37 (s), 0.85 (s) and 1:06 (d, J = 6 Hz);
and one methyl group for C 25 at δ 0.66 ppm (d, J = 6 Hz); an ethylene group at δ 5:51 ppm (br d, J = 5.7 Hz); signals of
the protons linked to C 26 at δ 4:13 and 3.49 ppm (each br d, J = 9.3 Hz and 9 Hz), and three anomeric protons at δ 6:43
ppm (br s); 5:56 ppm (br s) and 4:57 ppm (d, J = 7.0 Hz). From the above the data it can be assumed that the isolate is a
spirostan steroidal glicoside.Keywords: isolation, identification, Cordyline terminalis Kunth, Liliaceae

PRELIMINARY saponins which is a major compound in terms of the

stability of the foam formed (Bogoriani, 2001) and the
Indonesia is one country that is rich in flora steroid saponin compounds with heavy spirostananol
and fauna which are biological resources. Therefore 7.5 mg based on data high performance liquid
every species of plants, animals and micro-organisms chromatography chromatogram with the structure of
found on land and in sea have the values of the the atom C 25 and C 27 is a double bond (elesometilin
chemical in a sense produce chemicals that much groups) having a molecular weight of 866 and this
amount, the biodiversity available in Indonesia can be class of compounds have toxic properties on shrimp
interpreted as a source for a variety of chemicals
larvae ( Artenia salina Lich) identified positively
beaneka (Blunden, et al., 1981).
correlated to the antitumor compounds (Bogoriani, et
Plant carriage ( Cordyline terminalis Kunth)
al., 2007). Saponins are a complex group of natural
is a herbaceous plant familia Liliaceae, which
compounds, which have the molecular era bi and
traditional it leaves are used as diarrhea and
usefulness large (Konoshima, et al., 1995; Nakanishi,
dysentery. Ethnobotany approach provides an
1974; Agrawal, 992; Burger, et al., 1998). In this
assumption that the carriage leaves are compound
research will be the isolation and identification of
active against diarrhea and dysentery, so we need
steroidal saponins from leaves another carriage. ISSN
more research (Heftmann, 1974; Lajis, 1985 Mahato,
1907-9850
et al. 1982; Hostettmann and Marston, 1995;
Silverstein, et al., 1991).
From the results of previous studies have
been published the first of the compound saponin leaf
carriage. Type saponin contains is steroidal

40
MATERIALS AND METHODS Materials
Chemicals used consists of various types of
organic solvents and pro technical analysis (pa),10%
solution of sulfuric acid in ethanol, Liebermann
Burchard reagent (sulfuric acid + acetic anhydride),
silica gel 60 F 254for thin-layer chromatography and
silica gel 60 (70-230 mesh) for column
chromatography gravity.
Equipment
The equipment used consisted of a variety of
glassware used in the Organic Chemistry Laboratory,
analytical balance, suction vacuum gasing Buchi R-
114 is equipped with a vacuum system Buchi B-169,
and various analytical instruments as proton magnetic
resonance spectrometry (RMI1 H), and mass
spectrometry. Procedure Dry powder carriage leaves
approximately 0.5 kg was extracted by maceration for
24 hours using a solvent n- hexane to extract lipid.
Next resedu dried at room temperature until free n-
hexane, weighed, then macerated with methanol in an
iterative manner until extractable perfect, then
evaporated.
Condensed methanol extract obtained
partitioned between water and n- butanol, then the
fraction n- butanol is evaporated, washed with
diethylether, dissolved in methanol, and filtered. The
filtrate is then added diethylether excess methanol.
The precipitate is filtered (Heftmann, 1974). The
precipitate was then separated and purified saponin.
The process of separation and purification is done by
thin layer chromatography, column chromatography
and high performance liquid chromatography.
Saponins and steroid screening method
follows the method developed by Webb (Lajis, 1985).
After the process of separation and purification, pure
isolates, next made structure elucidation techniques
spectrometry.
41
RESULTS AND DISCUSSION
Extraction
A total of 0.5 kg of dry powder Andong leaves
extracted by maceration technique, successively using
solvent n- hexane and methanol. Solvent extraction
process with these two done to separate all the
components of both polar and non-polar of footage, thus
simplifying the subsequent separation. Condensed extract
of methanol is 85 g. Results of phytochemical screening
of the condensed methanol extract obtained that Andong
leaves contain saponins and steroids.
Fractionation
The methanol extract thick weighing 60 g and then
partitioned between water and n- butanol (1: 1), then each
fraction is separated and concentrated to obtain the water
fraction, and nbutanol. Each faction thick n-butanol
obtained weighing 40.1 g and 16.0 g of water. Fraction
nbutanol condensed contains more saponin after
phytochemical test. Fraction n- butanol then washed with
ether, After that dissolved in methanol, filtered and
then filtrate excess methanol added ether and the
precipitate is filtered. Saponin obtained precipitate weighing
11.0 g.
Separation and Purification
Fraction n- The most active butanol is then
separated by gravity column chromatography and high
performance liquid chromatography. Fraction n- butanol
total (Three grams) separated on a column using the
stationary phase silica gel 60 (70-230 mesh) and a mobile
phase of
chloroform-methanol-water (3: 1: 0.1) and stain sulfuric acid
10%). Results gravity column chromatography was 50
fractions (each fraction three meliliter). After being treated
chromatography thin-layer kloroformmethanol-water (3: 1:
0.1) eluate fractions produced three groups. Fraction
fraction B showed a stain through the test of purity by TLC
using various eluent. From the results of the foam test and
steroids showed that the fraction B is positive steroid
saponin.Analysis of B followed by HPLC fractions is by YMC
ODS-AQ column 5 μ m 120 A o 250 x 4.6 mm, the mobile
phase mixture of acetonitrile-water-acid acetate (50: 50:
0.05); show there five peaks later separation and
purification. The four components are separated
successively gained weight (4.8 mg, 1.4 mg; 4.0; 7.5 mg and
4.8 mg). Major isolates (7.5 mg) in the form of a white
powder which is forwarded for analysis by mass
spectrometry (MS) and proton magnetic resonance (RMI- 1
H).
Data Mass Spectrometry (MS)
From the calculation of the peak ion at m / z
891 [M + Na] + and 869 [M + H] + in the mass
spectrum shows that the isolates CT-4 has a
molecular weight of 868. Based on data from mass
spectrometry spectrum of isolates that have the price
of m / z 891 [M + Na] +, 869 [M + H] + and results of
data fragments fragments on the price of m / z 727 [(M
+ Na) - 164] +, 723 [(M + H) - 146] +, 705 [(M + H) -
JOURNAL OF CHEMICAL 2 (1), JANUARY 2008: 40-44
164] +, and 413 [(M + H) - 456 ] + indicates that three
sugar molecules bind isolates (possibility of two sugar
terminal and a central sugar) from metilpentosa
molecular weight of each sugar is 164 attached to the
aglycone.
Proton Magnetic Resonance Spectrometry Data
(RMI- 1 H)
Proton magnetic resonance spectrum in solvent C 5
D 5 N (300 MHz) showed signals of protons of four
methyl steroid at: δ 1.37 ppm (3H, s, 19-H), 1.06 ppm
(3H, d, J = 6 Hz, 21-H) and δ 0.85 ppm (3H, s, 18-H),
and a methyl group attached to the C 25 with δ 0.66
ppm (doublet, J = 6 Hz), an ethylene group in δ 5.51
ppm (1H, br d, J = 5.7 Hz, 6-H) and emerging signals
of protons bound to the atom C 26 on δ 4.13 and 3.49
ppm (each 1H, brd, J = 9.3 Hz, and 9HZ 26a and 26b-
H-H) and the three signals from the anomeric protons
δ 6.43 ppm (1H, br s, 1H), 5.56 ppm (1H, br s, 1H)
and 4.57 ppm (1H, d, J = 7.0 Hz, 1 H) reinforce the
notion that the compound is a steroidal saponins
derived isolates spirostan by tying three sugars
(allegedly two Research previous be produced
saponins steroidal saponins spirostananol structure
at the atomic C 25 and C 27 is a double bond
(elesometilin groups) having a molecular weight of
866 whereas the steroid saponin in this study had a
molecular weight of 868 and the C atom 25 tie the
methyl group derived from C 27.
CONCLUSIONS AND
RECOMMENDATIONS Conclusions
Based on the results research obtained can
be summarized as follows:
42
1. It has been done the isolation and identification
spirostan steroidal glycoside from leave Andong
( Cordyline terminalis Kunth) in the form of a
white amorphous solid.
2. Part sugar saponin showed three sugar bound to
aglikonnya. From the release of fragments of
146, 164 and 456 are from metilpentosa. of price
constant third anomeric proton coupling sugar
has three bond orientation ie two α-L-
ramnopiranosida and one β- D-fukopiranosida
that occurs either between or among glikon
glikon and sapogenin.
3. The structure of steroid sapogenin spirostan and
three sugars that make up glikon(two α- L-
ramnopiranosidadan one β- D- fukopiranosida).
Suggestion
From the results obtained, it may be advisable
1. To determine the structure of steroidal saponins from
Andong leaf (Cordyline terminalis Kunth) completely
necessary to measure nuclear magnetic resonance
spectrometry cutting-edge techniques.
2. To determine the type of sugar that make up
glikonnya must be done to isolate hydrolysis
technique. Synthesis technique needs to be done
and "modeling" computers to establish the proposed
structure as the absolute structure.
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